Integrated molecular and bioprocess engineering for bacterially produced immunogenic modular virus‐like particle vaccine displaying 18 kDa rotavirus antigen. Issue 2 (17th August 2016)

• Record Type: Journal Article
• Authors: Tekewe, Alemu; Fan, Yuanyuan; Tan, Emilyn; Middelberg, Anton P.J.; Lua, Linda H.L.
• Is Part Of: Biotechnology and bioengineering. Volume 114:Issue 2(2017)
• Page Start: 397
• Page End: 406
• Publication Date: 2016-08-17
• Subjects: capsomeres -- co‐expression -- selective precipitation -- murine polyomavirus -- large antigen -- Escherichia coli; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.26068
• Languages: English
• ISSNs: 0006-3592

Abstract

ABSTRACT: A high global burden of rotavirus disease and the unresolved challenges with the marketed rotavirus vaccines, particularly in the developing world, have ignited efforts to develop virus‐like particle (VLP) vaccines for rotavirus. While rotavirus‐like particles comprising multiple viral proteins can be difficult to process, modular VLPs presenting rotavirus antigenic modules are promising alternatives in reducing process complexity and cost. In this study, integrated molecular and bioprocess engineering approaches were used to simplify the production of modular murine polyomavirus capsomeres and VLPs presenting a rotavirus 18 kDa VP8* antigen. A single construct was generated for dual expression of non‐tagged murine polyomavirus capsid protein VP1 and modular VP1 inserted with VP8*, for co‐expression in Escherichia coli . Co‐expressed proteins assembled into pentameric capsomeres in E. coli . A selective salting‐out precipitation and a polishing size exclusion chromatography step allowed the recovery of stable modular capsomeres from cell lysates at high purity, and modular capsomeres were successfully translated into modular VLPs when assembled in vitro. Immunogenicity study in mice showed that modular capsomeres and VLPs induced high levels of VP8*‐specific antibodies. Our results demonstrate that a multipronged synthetic biology approach combining molecular and bioprocess engineering enabled simple and low‐cost production of highly immunogenic modular capsomeres and VLPs presenting conformational VP8* antigenic modules. This strategy potentially provides a cost‐effective production route for modular capsomere and VLP vaccines against rotavirus, highly suitable to manufacturing economics for the developing world. Biotechnol. Bioeng. 2017;114: 397–406. © 2016 Wiley Periodicals, Inc. Abstract : A multipronged approach for simplified production of highly immunogenic modular virus‐like particles displaying 18 kDa rotavirus antigens. Integrating Escherichia coli protein co‐expression, non‐chromatography purification and cell‐free in vitro virus‐like particle assembly resulted in a highly stable bacterial‐based modular virus‐like particles displaying conformational rotavirus VP8* antigenic modules. These modular virus‐like particles induced high titres of VP8* antigen‐specific antibodies in mice.